原发性局灶节段性肾小球硬化ACTN4和SYNPO基因启动子的突变  

Mutations of ACTN4 and SYNPO genes promoter in primary focal segmental glomerulosclerosis

作　　者：戴胜川[1] 王朝晖[1] 潘晓霞[1] 王伟铭[1] 张文[1] 任红[1] 谢静远[1] 朱斌[1] 冯旗[2] 陈楠[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾脏科,200025 [2]中国科学院上海生命科学院国家基因中心

出　　处：《中华肾脏病杂志》2009年第8期613-618,共6页Chinese Journal of Nephrology

基　　金：上海市卫生局重点学科基金（05111001）;上海市重点学科基金（T0201）;上海市领军人才基金;上海市科委重点项目（07JC14037）

摘　　要：目的探讨散发性原发性局灶节段性肾小球硬化（FSGS）患者中ACTN4和SYNPO基因启动子区突变的致病作用。方法盐析法提取82例FSGS患者外周血基因组DNA，经引物设计及PCR扩增后测序。突变位点经转录因子结合模拟软件筛选，pGL3-Basic构建表达载体与pRL—SV40质粒瞬时共转染PCI2细胞，双荧光素酶法检测基因表达。检测患者父母头发DNA。免疫荧光检测患者肾组织α-actinin-4和synaptopodin蛋白表达。结果3例ACTN4基因突变分别为1—34C〉T、1～590delA和（1—1044delT）＋（1—797T〉C）＋（1—769A〉G）。2例SYNPO基因突变分别为1—24G〉A和1—851C〉T。1例患者分别接受父母1—1044delT和1—797T〉C变异。除1—1044delT组外，变异组荧光素酶表达强度比正常组有不同程度下降，与突变患者肾组织α-actinin-4和synaptopodin免疫荧光强度下降基本相符。结论ACTN4和SYNPO基因启动子区顺式作用元件区的变异影响基因的转录，可能在散发性FSGS发病中起作用。Objective To investigate the mutations ACTN4 and SYNPO genes promoter in sporadic primary focal segmental glomerulosclerosis （FSGS） and to analyze the role of mutations in FSGS. Methods The study consisted of 82 Chinese primary FSGS, including 39 females and 43 males, ranged from 12 to 76 years old. Seventy volunteers were selected as heahhy control group. Genomic DNA was extracted from peripheral blood cells of FSGS patients and hair of patients＇ parents by polymerase chain reaction （PCR） and direct sequencing to analyze ACTN4 and SYNPO gene promoter mutations. Mutations were matched with GenBank and TRANSFAC software database （www.ncbi.nlm.nih.gov; www.genometix.de; www.gene-regulation, corn）. Dual luciferase assay system was used to analyze the promoter region mutations, based on PGL3-Basic vector, pRL- SV40 and PC12 cell line. Hair DNA of novel mutation patients＇ parents was sequenced. Expression of alpha-actinin-4 and synaptopodin in patients ＇ kidney tissue was examined by

关 键 词：肾小球硬化症 局灶节段性 辅肌动蛋白 启动区(遗传学) 突变 ACTN4 SYNPO 

分 类 号：R692.6[医药卫生—泌尿科学] R562.25[医药卫生—外科学]